Comparative Analysis of the Proteome in the Peel and Flesh of ‘Hongro’ Apples

Jung-Sung Chung; Sung Hwan Choi; Jae Ho Kim; Soo Yong Shim; Moch R. Adnan; Eun-Sil Chang; Young-Geol Sohn; Yun-Hee Kim; Jin Gook Kim; Jeung Joo Lee

doi:10.7235/HORT.20210017

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2021 Vol.39, Issue 2 Preview Page Next Page

Research Article

Comparative Analysis of the Proteome in the Peel and Flesh of ‘Hongro’ Apples ‘홍로’ 사과 과피 및 과육의 단백질체 비교 분석

30 April 2021. pp. 191-203

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Abstract

The proteome expressed in the peel and flesh of ‘Hongro’, a domestic breeding apple cultivar, was extracted and separated by two-dimensional electrophoresis. Most of the proteins were expressed at similar levels in the peel and flesh, but 30 protein spots showed a difference of more than two times higher in expression level in the peel and flesh. Of these, 25 were higher in the peel and 5 were higher in the flesh. By analyzing these proteins with MALDI-TOF/TOF, 18 of the protein spots that were higher in the peel were identified as proteins with their respective functions, and the remaining 7 were identified as having unknown function or were unidentified. In the case of the protein spots expressed higher in the flesh, two were identified as proteins having respective functions, and the other three were unidentified. Proteins more highly expressed in the peel were 1-aminocyclopropane- 1-carboxylate oxidase homolog (spot 21) related to the biosynthesis of ethylene; acetyl-CoA acetyltransferase (spot 148), cytosolic 1, acetyl-CoA acetyltransferase, cytosolic 1-like (spot 316) and 8-hydroxygeraniol dehydrogenase-like (spot 348) related to the biosynthesis of secondary metabolites; oxygen-evolving enhancer protein 1, chloroplastic (spot 210), and ribulose-1, 5-bisphosphate carboxylase/oxygenase large subunit (spot 273) related to the photosynthesis; (+)-neomenthol dehydrogenase-like (spot 216 and 219), formate dehydrogenase, mitochondrial (spot 305), glutathione S-transferase-like (spot 483 and 484), and acidic endochitinase SE 2-like, partial (spot 391 and 392) related to the biological defense and stress response; and antioxidant enzyme L-ascorbate peroxidase 2 (spot 221), MLP-like protein 34 (spot 406), major allergen Mal d 1 (spot 451 and 452), and major allergen Mal d 1.06A01 (spot 457) related to allergies. On the other hand, proteins that were more highly expressed in the flesh were chaperonin 60 subunit alpha 1, chloroplastic (spot 8) related to protein folding, and NADP-dependent D-sorbitol-6-phosphate dehydrogenase (spot 380) related to carbohydrate metabolism. These results will be useful in a wide range of studies related to apple breeding, storage, quality, and environmental tolerance.

Keywords

Allergy protein

biological defense and stress protein

ethylene biosynthetic protein

Malus domestica

proteomics

secondary metabolic protein

국내 육종 사과 품종인 ‘홍로’의 과피와 과육에서 발현되는 단백질체를 추출하여 이차원 전기영동으로 분리하였다. 그 결과 대부분의 단백질들은 과피와 과육에서 유사한 정도로 발현되었으나, 30개의 단백질 spot들은 과피 또는 과육에서의 발현 정도가 2배 이상 차이가 났다. 이들 중 25개는 과피에서, 5개는 과육에서 더 높게 발현되었다. 이들 단백질들을 MALDI-TOF/TOF로 분석한 결과, 과피에서 더 높게 발현된 단백질 spot들 중 18개는 각각의 기능을 갖는 단백질로 동정 되었고, 나머지 7개는 기능이 밝혀지지 않은 것으로 동정되었거나 미동정 되었다. 과육에서 더 높게 발현된 단백질 spot들의 경우 2개는 각각의 기능을 갖는 단백질로 동정 되었고, 나머지 3개는 미동정 되었다. 과피에서 더 높게 발현된 단백질은 에틸렌의 생합성과 관련된 1-aminocyclopropane-1-carboxylate oxidase homolog(spot 21번), 이차대사산물의 생합성과 관련된 acetyl-CoA acetyltransferase, cytosolic 1(spot 148), acetyl-CoA acetyltransferase, cytosolic 1-like(spot 316) 및 8-hydroxygeraniol dehydrogenase-like (spot 348), 광합성과 관련된 oxygen-evolving enhancer protein 1, chloroplastic(spot 210) 및 ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit(spot 273), 생체방어 및 스트레스 반응과 관련된 (+)-neomenthol dehydrogenase-like (spot 216 및 219), formate dehydrogenase, mitochondrial(spot 305), glutathione S-transferase-like(spot 483 및 484) 및 acidic endochitinase SE 2-like, partial(spot 391 및 392), 항산화효소인 L-ascorbate peroxidase 2(spot 221), 알레르기 관련 단백질인 MLP-like protein 34(spot 406), major allergen Mal d 1(spot 451 및 452) 및 major allergen Mal d 1.06A01(spot 457)이었다. 한편 과육에서 더 높게 발현된 단백질은 protein folding과 관련된 chaperonin 60 subunit alpha 1, chloroplastic (spot 8) 및 탄수화물 대사와 관련된 NADP-dependent D-sorbitol-6-phosphate dehydrogenase(spot 380)이었다. 이상의 연구 결과들은 사과의 육종, 저장, 품질, 환경에 대한 내성 등과 관련된 광범위한 연구에 유용하게 이용될 수 있을 것으로 사료되었다.

키워드

알레르기 단백질

생체방어 및 스트레스 단백질

에틸렌 생합성 단백질

Malus domestica

프로테오믹스

이차대사 단백질

References

Alexander L, Grierson D (2002) Ethylene biosynthesis and action in tomato: a model for climacteric fruit ripening. J Exp Bot 53:2039-2055. doi:10.1093/jxb/erf072 10.1093/jxb/erf07212324528

Aprea E, Charles M, Endrizzi I, Corollaro ML, Betta E, Biasioli F, Gasperi F (2017) Sweet taste in apple: the role of sorbitol, individual sugars, organic acids and volatile compounds. Sci Rep 7:44950. doi:10.1038/srep44950 10.1038/srep4495028322320PMC5359574

Archetti M, Döring TF, Hagen SB, Hughes NM, Leather SR, Lee DW, Lev-Yadun S, Menetas Y, Ougham HJ, et al. (2009) Trends Ecol Evol 24:166-173. doi:10.1016/j.tree.2008.10.006 10.1016/j.tree.2008.10.00619178979

Armstrong GA, Hearst JE (1996) Genetics and molecular biology of carotenoid pigment biosynthesis. FASEB J 10:228-237. doi:10.1096/fasebj.10.2.8641556 10.1096/fasebj.10.2.86415568641556

Blanco C, Diaz-Perales A, Collada C, Sanchez-Monge R, Aragoncillo C, Castillo R, Ortega N, Alvarez M, Carrilo T, et al. (1999) Class I chitinases as potentioal panallergens involved in the latex-fruit syndrome. J Allergy Clin Immunol 103:507-513. doi:10.1016/S0091-6749(99)70478-1 10.1016/S0091-6749(99)70478-1

Bohlmann J, Keeling CI (2008) Teropenoid biomaterials. Plant J 54:656-669. doi:10.1111/j.1365-313X.2008.03449.x 10.1111/j.1365-313X.2008.03449.x18476870

Boyer J, Liu RH (2004) Apple phytochemicals and their health benefits. Nutr J 3:1-15. doi:10.1186/1475-2891-3-5 10.1186/1475-2891-3-515140261PMC442131

Bradford MM (1976) A rapid sensitive methods for the quantization of microgram quantities of protein utilizing the principles of protein-dye binding. Anal Biochem 72:248-255. doi:10.1016/0003-2697(76)90527-3 10.1016/0003-2697(76)90527-3

Brehler R, Theissen U, Mohr C, Luger T (1997) Latex-fruit syndrome: frequency of cross-reacting IgE antibodies. Allergy 52:404-410. doi:10.1111/j.1398-9995.1997.tb01019.x 10.1111/j.1398-9995.1997.tb01019.x9188921

Choi HW, Lee BG, Kim NH, Park Y, Lim CW, Song HK, Hwang BK (2008) A role for a menthone reductase in resistance against microbial pathogens in plants. Plant Physiol 148:383-401. doi:10.1104/pp.108.119461 10.1104/pp.108.11946118599651PMC2528125

Christopher MR, Gayle MV, Ann AR, Adam DH, Dale RL, Patrick AR, Philip LF (2009) Genetic diversity and population structure in Malus sieversii, a wild progenitor species of domesticated apple. Tree Genet Genomes 5:339-347. doi:10.1007/s11295-008-0190-9 10.1007/s11295-008-0190-9

Davies KM (2004) An introduction to plant pigments in biology and commerce. In KM Davies, ed, Plant pigments and their manipulation: Annual Plant Reviews, 14. Blackwell Publishing, Oxford, UK, pp 1-18. doi:10.1002/9781119312994.apr0131 10.1002/9781119312994.apr0131

Davletova S, Rizhsky L, Liang H, Shengqiang Z, Oliver DJ, Coutu J, Shulaev V, Schlauch K, Mittler R (2005) Cytosolic ascorbate peroxidase 1 is a central component of the reactive oxygen gene network of Arabidopsis. Plant Cell 17:268-281. doi:10.1105/tpc.104.026971 10.1105/tpc.104.02697115608336PMC544504

Douglas KT (1987) Mechanism of action of glutathione-dependent enzymes. Adv Enzymol Relat Areas Mol Biol 59:103-167. doi:10.1002/9780470123058.ch3 10.1002/9780470123058.ch3

Elzebroek ATG, Wind K (2008) Apple. In ATG Elzebrock and K Wind, ed, Guide to cultivated plants. CAB International, Wallingford, UK, pp 25-28 10.1079/9781845933562.0000

Fan X, Blankenship SM, Mattheis JP (1999) 1-Methylcyclopropene inhibits apple ripening. J Am Soc Hortic Sci 124:690-695. doi:10.21273/JASHS.124.6.690 10.21273/JASHS.124.6.690

Finkelstein R (2013) Abscisic Acid Synthesis and Response. The Arabidopsis Book 11:e0166. doi:10.1199/tab.0166 10.1199/tab.016624273463PMC3833200

Guarino C, Arena S, De Simone L, D'Ambrosio C, Santoro S, Rocco M, Scaloni A, Marra M (2007) Proteomic analysis of the major soluble components in Annurca apple flesh. Mol Nutr Food Res 51:255-262. doi:10.1002/mnfr.200600133 10.1002/mnfr.20060013317266180

Hartl FU, Hayer-Hartl M (2009) Converging concepts of protein folding in vitro and in vivo. Nat Struct Mol Biol 16:574-581. doi:10.1038/nsmb.1591 10.1038/nsmb.159119491934

Herndl A, Marzban G, Hahn R, Boscia D, Hemmer W, Maghuly F, Stoyanova E, Katinger H (2007) Mapping of Malus domestica allergens by 2-D electrophoresis and IgE-reacivity. Electrophor 28:437-438. doi:10.1002/elps.200600342 10.1002/elps.20060034217195260

Holman JD, Dasari S, Tabb DL (2013) Informatics of protein and posttranslational modification detection via shotgun proteomics. Methods Mol Biol 1002:167-179. doi:10.1007/978-1-62703-360-2_14 10.1007/978-1-62703-360-2_1423625403PMC4012295

Hourton-Cabassa C, Ambard-Bretteville F, Moreau F, Davy de Virville J, Remy R, Colas des Francs-Small C (1998) Stress induction of mitochondrial formate dehydrogenase in potato leaves. Plant Physiol 116:627-635. doi:10.1104/pp.116.2.627 10.1104/pp.116.2.6279490763PMC35120

Hurkman WJ, Tanaka CK (1986) Solubilization of plant membrane proteins for analysis by two-dimensional gel glectrophoresis. Plant Physiol 81:802-806. doi:10.1104/pp.81.3.802 10.1104/pp.81.3.80216664906PMC1075430

Ikeda H, Esaki N, Nakai S, Hashimoto K, Uesato S, Soda K, Fujita T (1991) Acyclic monoterpene primary alcohol:NADP⁺ oxidoreductase of Rauwolfia serpentina cells: the key enzyme in biosynthesis of monoterpene alcohols. J Biochem 109:341-347

Izumi M, Tsunoda H, Suzuki Y, Makino A, Ishida H (2012) RBCS1A and RBCS3B, two major members within the Arabidopsis RBCS multigene family, function to yield sufficient Rubisco content for leaf photosynthetic capacity. J Exp Bot 63:2159-2170. doi:10.1093/jxb/err434 10.1093/jxb/err43422223809PMC3295403

Jetter R, Kunst L, Samuels AL (2006) Composition of plant cuticular waxes. In M Riederer, C Muller, eds, Biology of the plant cuticle, Blackwell, Oxford, UK, pp 145-181. doi:10.1002/9780470988718.ch4 10.1002/9780470988718.ch4

Kanayama Y, Mori H, Imaseki H, Yamaki S (1992) Nucleotide sequence of a cDNA encoding NADP-sorbitol-6-phosphate dehydrogenase from apple. Plant Physiol 100:1607-1608. doi:10.1104/pp.100.3.1607 10.1104/pp.100.3.160716653170PMC1075832

Kim JH, Shim SY, Chang ES, Sohn YG, Kim YH, Kim JG, Lee JJ (2020) A Large-scale proteome analysis of proteins expressed in the peel of the Malusdomestica 'Hong-Ro' apple. Hortic Sci Technol 38:795-809

Kolattukudy PE (2001) Polyesters in higher plants. Adv Biochem Eng Biot 71:1-49. doi:10.1007/3-540-40021-4_1 10.1007/3-540-40021-4_111217409

Kwon YS, Lee DY, Rakwal R, Baek SB, Lee JH, Kwak YS, Seo JS, Chung WS, Bae DW, et al. (2016) Proteomic analyses of the interaction between the plant-growth promoting rhizobacterium aenibacillus polymyxa E681 and Arabidopsis thaliana. Proteom 16:122-135. doi:10.1002/pmic.201500196 10.1002/pmic.20150019626460066

Lattanzio V, Lattanzio VMT, Cardinali A (2006) Role of phenolics in the resistance mechanisms of plants against fungal pathogens and insects. In F Imperato, ed, Phytochemistry: Advances in Research, Research Singpost, Kerala, India, pp 23-67

Leaver MJ, George SG (1998) A piscine glutathione S-transferase which efficiently conjugates the end-roducts of lipid peroxidation. Mar Environ Res 46:71-74. doi:10.1016/S0141-1136(97)00071-8 10.1016/S0141-1136(97)00071-8

Lee JJ, Park KW, Kwak YS, Ahn JY, Jung YH, Lee BH, Jeng JC, Lee HS, Kwak SS (2012) Comparative proteomic study between tuberous roots of light orange- and purple-fleshed sweetpotato cultivars. Plant Sci 193-194:120-129. doi:10.1016/j.plantsci.2012.06.003 10.1016/j.plantsci.2012.06.00322794925

Marondedze C, Thomas LA (2012) Apple Hypanthium Firmness: New Insights from Comparative Proteomics. Appl Biochem Biotechnol 168:306-326. doi:10.1007/s12010-012-9774-9 10.1007/s12010-012-9774-922733236

Martin LBB, Rose JKC (2014) There's more than one way to skin a fruit: formation and functions of fruit cuticles. J Exp Bot 65:4639-4651. doi:10.1093/jxb/eru301 10.1093/jxb/eru30125028557

Marzban G, Puehringer H, Dey R, Brynda S, Ma Y, Martinelli A, Zaccarini M, van der Weg E, Housley Z, et al. (2005) Localisation and distribution of the major allergens in apple fruits. Plant Sci 169:387-394. doi:10.1016/j.plantsci.2005.03.027 10.1016/j.plantsci.2005.03.027

Matsui NM, Smith-Beckerman DM, Epstein LB (1999) Staining of preparative 2-D gels. In AJ Link, ed, 2-D Proteome Analysis Protocols, Humana Press, Totowa, New Jersey, USA, pp 301-311

Meriläinen G, Poikela V, Kursula P, Wierenga RK (2009) The thiolase reaction mechanism: the importance of Asn316 and His348 for stabilizing the enolate intermediate of the Claisen condensation. Biochem 48:11011-11025. doi:10.1021/bi901069h 10.1021/bi901069h19842716

Palma JM, Corpas FJ, del Río LA (2011) Proteomics as an approach to the understanding of the molecular physiology of fruit development and ripening. J Proteom 74:1230-1243. doi:10.1016/j.jprot.2011.04.010 10.1016/j.jprot.2011.04.01021524723

Pirrung MC (1999) Ethylene biosynthesis from 1-aminocyclopropanecarboxylic acid. Acc Chem Res 32:711-718. doi:10.1021/ar960003 10.1021/ar960003

Popov VO, Lamzin VS (1994) NAD (+)-dependent formate dehydrogenase. Biochem J 301:625-643. doi:10.1042/bj3010625 10.1042/bj30106258053888PMC1137035

Pühringer H, Moll D, Hoffmann-Sommergruber K, Watillon B, Katinger H, Laimer da Câmara Machado M (2000) The promoter of an apple YPR10 gene, encoding the major apple allergen Mal d 1, is stress and pathogen-inducible. Plant Sci 152:35-50. doi:10.1016/S0168-9452(99)00222-8 10.1016/S0168-9452(99)00222-8

Qin G, Wang Q, Liu J, Li B, Tian S (2009) Proteomic analysis of changes in mitochondrial protein expression during fruit senescence. Proteom 9:4241-4253. doi:10.1002/pmic.200900133 10.1002/pmic.20090013319688753

Quideau S, Deffieux D, Douat-Casassus C, Pouységu (2011) Plant polyphenols: Chemical properties, biological activities, and synthesis. Angew Chem 50:586-621. doi:10.1002/anie.201000044 10.1002/anie.20100004421226137

Ribeiro FA, Gomes de Moura CF, Aguiar O Jr, de Oliveira F, Spadari RC, Oliveira NR, Oshima CT, Ribeiro DA (2014) The chemopreventive activity of apple against carcinogenesis: antioxidant activity and cell cycle control. Eur J Cancer Prev 23:477-480. doi:10.1097/CEJ.0000000000000005 10.1097/CEJ.000000000000000524366437

Salcedo G, Sanchez-Monge R, Diaz-Perales A, Garcia-Casado G, Barber D (2004) Plant non-specific lipid transfer proteins as food and pollen allergens. Clin Exp Allergy 34:1336-1341. doi:10.1111/j.1365-2222.2004.02018.x 10.1111/j.1365-2222.2004.02018.x15347364

Sancho AI, Foxall R, Rigby NM, Browne T, Zuidmeer L, van Ree R, Waldron KW, Mills EN (2006a) Maturity and storage influence on the apple (Malus domestica) allergen Mal d 3, a nonspecific lipid transfer protein. J Agric Food Chem 54:5098-5104. doi:10.1021/jf0530446 10.1021/jf053044616819922

Sancho AI, Foxall R, Browne T, Dey R, Zuidmeer L, Marzban G, Waldron, KW, van Ree R, et al. (2006b) Effect of postharvest storage on the expression of the apple allergen Mal d 1. J Agric Food Chem 54:5917-5923. doi:10.1021/jf060880m 10.1021/jf060880m16881695

Shin MH, Muneer S, Kim YH, Lee JJ, Bae DW, Kwack YB, Kumarihami HMPC, Kim JG (2020) Proteomic analysis reveals dynamic regulation of fruit ripening in response to exogenous ethylene in kiwifruit cultivars. Hortic Environ Biotechnol 61:93-114. doi:10.1007/s13580-019-00209-6 10.1007/s13580-019-00209-6

Suzuki K, Nakanishi H, Bower J, Yoder DW, Osteryoung KW, Miyagishima SY (2009) Plastid chaperonin proteins Cpn60 alpha and Cpn60 beta are required for plastid division in Arabidopsis thaliana. BMC Plant Biol 9:38-38. doi:10.1186/1471-2229-9-38 10.1186/1471-2229-9-3819344532PMC2670834

Teo G, Suzuki Y, Uratsu SL, Lampinen B, Ormonde N, Hu WK, DeJong TM, Dandekar AM (2006) Silencing leaf sorbitol synthesis alters long-distance partitioning and apple fruit quality. PNAS 103:18842-18847. doi:10.1073/pnas.0605873103 10.1073/pnas.060587310317132742PMC1693749

van Ree R (2002) Clinical importance of non-specific lipid transfer proteins as food allergens. Biochem Soc Trans 30:910-913. doi:10.1042/bst0300910 10.1042/bst030091012440944

Velasco R, Zharkikh A, Affourtit J, Dhingra A, Cestaro A, Kalyanaraman A, Fontana P, Bhatnagar SK, Troggio M, et al. (2010) The genome of the domesticated apple (Malus × domestica Borkh.). Nat Genet 42:833-841. doi:10.1038/ng.654 10.1038/ng.65420802477

Vershinin A (2008) Biological functions of carotenoids ‐ diversity and evolution. Biofactors 10:99-104. doi:10.1002/biof.5520100203 10.1002/biof.552010020310609869

Yang H, Zhang T, Masuda T, Lv C, Sun L, Qu G, Zhao G (2011) Chitinase III in pomegranate seeds (Punica granatum Linn): a high-capacity calcium-binding protein in amyloplasts. Plant J 68:765-776. doi:10.1111/j.1365-313X.2011.04727.x 10.1111/j.1365-313X.2011.04727.x21790816

Yi X, McChargue M, Laborde S, Frankel LK, Bricker TM (2005) The manganese-stabilizing protein is required for photosystem II assembly/stability and photoautotrophy in higher plants. J Biol Chem 280:16170-16174. doi:10.1074/jbc.M501550200 10.1074/jbc.M50155020015722336

Zheng Q, Song J, Campbell-Palmer L, Thompson K, Li L., Walker B, Cui Y, Li X (2013) A proteomic investigation of apple fruit during ripening and in response to ethylene treatment. J Proteom 93:276-294. doi:10.1016/j.jprot.2013.02.006 10.1016/j.jprot.2013.02.00623435059

Information

Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
Publisher(Ko) :원예과학기술지
Journal Title :Horticultural Science and Technology
Journal Title(Ko) :원예과학기술지
Volume : 39
No :2
Pages :191-203
Received Date : 2020-10-12
Revised Date : 2020-11-05
Accepted Date : 2020-12-03
DOI :https://doi.org/10.7235/HORT.20210017

Horticultural Science and TechnologyISSN:1226-8763(Print) 2465-8588(Online)원예과학기술지

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